Article Figures & Data
Figures
- FIG. 1.
Map of the 19-kb region on the p3iANG plasmid, clustering together the three integrons and chloramphenicol, kanamycin, and tetracycline resistance genes (approximate sizes; not to scale). The upper and lower lines represent the amplicons obtained by the primer pairs as indicated and oriented by arrows. Primer pairs and resulting amplicons for sequencing the 3.30-kb amplicon are described in the text; the 140-bp junction between the two integrons is indicated by the solid black rectangle. Sequence DQ227350 corresponds to the 3.3-kb 3′CSrev/5′CSrev amplicon. Sequence DQ149925 corresponds to the contiguous 1.5-kb In-F/In-B amplicon.
Tables
- TABLE 1.
Vibrio strains under study
Strain Date of isolation (day/mo/yr) Place of isolation (Angolan province) Resistances in addition to common profilea V. cholerae O1 582 04/03/92 Luanda KAN, SUL, TMP, TET V. cholerae O1 583 09/03/92 Luanda KAN, SUL, TMP, TET V. cholerae O1 588 10/03/92 Luanda ERY, KAN, SUL, TMP, TET V. cholerae O1 611 26/03/92 Luanda KAN, SUL, TMP, TET V. cholerae O1 647 21/04/92 Luanda KAN, SUL, TMP, TET V. cholerae O1 817 28/06/94 Bengo (Caxito) KAN, SUL, TMP, TET V. cholerae O1 819 28/06/94 Bengo (Dondo) KAN, SUL, TMP, TET V. cholerae O1 1383 09/11/94 Benguela KAN, SUL, TMP, TET V. cholerae O1 1356 23/11/94 Cuando Cubango KAN, SUL, TMP, TET V. cholerae O1 1361 03/05/95 Cabinda KAN, SUL, TMP, TET V. cholerae O1 908 16/04/96 Cabinda ERY, KAN, SUL, TMP, TET V. parahaemolyticus 570 06/12/91 Luanda SUL, TMP V. parahaemolyticus 586 17/03/92 Luanda SUL, TMP V. cholerae non-O1 698b 02/03/93 Bengo River ERY, SUL, TET V. cholerae O1 547b 31/09/94 Bengo River KAN, SUL, TMP, TET ↵ a Abbreviations: AMP, ampicillin; CHL, chloramphenicol; ERY, erythromycin; KAN, kanamycin; PEN, penicillin; STR, streptomycin; SPT, spectinomycin; SUL, sulfamethoxazole; TET, tetracycline; TMP, trimethoprim. The common profile is resistance to AMP, CHL, PEN, STR, and SPT.
↵ b Environmental isolate.
- TABLE 2.
PCR primers used in this study
Primer and purpose Sequence (5′ to 3′) Target gene or description Amplicon size (bp) Reference, accession no., and/or chromosomal locationd Class 1 integron and cassette detection In-F GGCATCCAAGCAGCAAG Class 1 integron Variable 23 In-B AAGCAGACTTGACCTGA Dfr-Ba TCACCTTCTGGCTCAATGTCG dfrA15 490 Z83311, bp 374-354 Aad-Ba GCCGACTACCTTGGTGATCTCG aadA1 1,476 AY103457, bp 786-765 blaP1-Ba CTGGTTCATTTCAGATAGCG blaP1 874 11 Integron cluster mapping and characterization 3′CSrev AGTCCAGTTCAGACGAA Integron spacers Variable U12338, bp 1433-1416 5′CSrev GAACGACGAACCTACGG U12338, bp 4814-4831 Dfr-Fb GGAGTTATCGGAAATGGCCCAG Integron spacers Variable Z83311, bp 37-58 Bla-Fb GGCAATCAGCGCTTCCCGTT AF221899, bp 334-353 Dfr-F2 TTAGCCAAGACTTCGTGT dfrA15-sul1 AB113114, bp 515-532 Sul1B GCAAGGCGGAAACCCGCGCC X12869, bp 1360-1341 Int1A AAAACCGCCACTGCGCCGTTA intI1 1,200 12 Int1B GAAGACGGCTGCACTGAACG Sul1Ac GCACGTGCTGTCGAACCTTCA AY0462763, bp 38844-38865 Aph F GGCAATCAGGTGCGACAAT kan 484 AY090559, bp 12978-12997 Aph R GTGACGACTGAATCCGGTGA AY090559, bp 13461-13441 CAT1-F GGCATTTCAGTCAGTTG cat1 584 V00622, bp 312-328 CAT1-B CCGCCCTGCCACTCATC V00622, bp 880-896 TET(G) 1 GCTCGGTGGTATCTCTGCTC tetG 468 25 TET(G) 2 AGCAACAGAATCGGGAACAC ICE detection and characterization Int1-F GCTGGATAGGTTAAGGGCGG SXT int 592 8 Int1-B CTCTATGGGCACTGTCCACATTG Nint-F TACCTACAGCAGGAACGGGC SXT int 258 AF099172, bp 255-274 Nint-B GCAGCACAGACACCAGACGT AF099172, bp 513-494 FLOR-F TTATCTCCCTGTCGTTCCAGCG floR 526 22 FLOR-2 CCTATGAGCACACGGGGAGC TMP-F TGGGTAAGACACTCGTCATGGG dfr18 389 19 TMP-B ACTGCCGTTTTCGATAATGTGG SUL2-F AGGGGGCAGATGTGATCGC sul2 625 AY034138, bp 16726-16706 SUL2-B TGTGCGGATGAAGTCAGCTCC 19 STRA-F TTGATGTGGTGTCCCGCAATGC strA 383 19 STRA-B CCAATCGCAGATAGAAGGCAA strB-F GGCACCCATAAGCGTACGCC strB 470 10 strB-R TGCCGAGCACGGCGACTACC dfr1-F CGAAGAATGGAGTTATCGGG dfrA1 372 22 dfr1-B TGCTGGGGATTTCAGGAAAG - TABLE 3.
Resistances transferred and class 1 integron cassettes identified in Vibrio strains under study
Strain Transferred resistances in addition to common profilea Class 1 integron cassette(s) V. cholerae O1 582d KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b,g V. cholerae O1 583d KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 611d KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 1383d KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 647 KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 817 KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 819 KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 1356 KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 1361 KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 588d ERY, KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. cholerae O1 908 ERY, KAN, STR, SPT, TET dfrA15, blaP1, qacH-aadA8b V. parahaemolyticus 570 dfrA15 c , g V. parahaemolyticus 586 dfrA15 c , g V. cholerae non-O1 698d NCe V. cholerae O1 547 NCf dfrA15, blaP1, qacH-aadA8b ↵ a Abbreviations: AMP, ampicillin; CHL, chloramphenicol; ERY, erythromycin; KAN, kanamycin; PEN, penicillin; STR, streptomycin; SPT, spectinomycin; SUL, sulfamethoxazole; TET, tetracycline; TMP, trimethoprim. The transferred common profile is resistance to AMP, CHL, SUL, and TMP.
↵ b Plasmid located.
↵ c Chromosome located.
↵ d SXT integrase positive.
↵ e NC, not conjugative.
↵ f AMP, CHL, and TMP resistance transferred by electroporation.
↵ g Confirmed by amplicon sequencing.
